CN111909092A

CN111909092A - Fluorine-containing substituted imidazole salt compound, preparation method thereof, medicinal composition and application thereof

Info

Publication number: CN111909092A
Application number: CN201910387619.5A
Authority: CN
Inventors: 邓贤明; 林圣彩; 张宸崧
Original assignee: Xiamen Huachuo Biomedical Technology Co ltd
Current assignee: Xiamen Huachuo Biomedical Technology Co ltd
Priority date: 2019-05-10
Filing date: 2019-05-10
Publication date: 2020-11-10
Also published as: WO2020228596A1; US20220242828A1; WO2020228596A8; JP2022532006A; AU2020274362A1; EP3967683A1; TW202104192A; CN113966325A; CA3140417A1; EP3967683A4; CN113966325B; KR20220004730A; AU2020274362B2

Abstract

The invention relates to a compound with the activity of activating 5 ' -adenosine monophosphate activated protein kinase (AMPK), a preparation method thereof, a pharmaceutical composition containing the compound, and application of the compound in preparing medicaments for reducing fatty acid synthesis, inhibiting triglyceride and cholesterol synthesis, preventing and/or treating obesity and II type diabetes, preventing and/or treating tumors, preventing and/or treating Parkinson's disease, preventing and/or treating Alzheimer's disease or prolonging the life of mammals:

Description

Fluorine-containing substituted imidazole salt compound, preparation method thereof, medicinal composition and application thereof

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a compound with an activity of activating 5 ' -adenosine monophosphate activated protein kinase (AMPK), a preparation method thereof, a medicinal composition containing the compound, and application of the compounds in preparing medicaments for reducing fatty acid synthesis, inhibiting triglyceride and cholesterol synthesis, preventing and/or treating obesity and type II diabetes, preventing and/or treating tumors, preventing and/or treating Parkinson's disease, preventing and/or treating Alzheimer's disease or prolonging the life of mammals.

Background

The 5' -adenosine monophosphate activated protein kinase (AMPK), is the most important molecule for sensing energy levels and regulating metabolic homeostasis in the body and cells. AMPK is a heterotrimer composed of three subunits, α, β and γ, and is activated by phosphorylation of its upstream kinase under physiological conditions when the Cell is in an energy deficient state, initiating a series of downstream reactions, including promotion of hydrolysis of triglycerides in adipocytes, promotion of liver uptake of fatty acids in blood and oxidation, inhibition of fatty acid and cholesterol production and triglyceride formation, promotion of lipid oxidation and glucose uptake and breakdown in muscle, inhibition of insulin secretion and glycogen synthesis in islet β cells, inhibition of protein synthesis and promotion of autophagy and ketone body production, etc. (Hardie DG, Ross FA, hawlsa. nat Rev Mol Cell biol.201213 (4): 251-62). The result of these effects is to increase the energy-producing metabolism in vivo and decrease the energy-consuming metabolism in vivo, thereby achieving the effect of maintaining energy homeostasis and ensuring normal vital activities of cells. Thus, AMPK activation can have many beneficial health effects. For example, AMPK expression or activity is strongly inhibited in a variety of tumor tissues, such as melanoma, breast, colon, and lung cancer tissues, which further breaks the original balance of anabolism and catabolism in these tissues, exacerbating tumor development (Shackelford DB, Shaw rj. nat Rev cancer. 20099 (8): 563-75). At the cellular level, there have been numerous studies showing that activation of AMPK can inhibit the anabolism of tumor cells, for example by inhibiting the mTORC1 complex, thereby preventing their proliferation (Inoki K, Kim J, Guan KL. Annu Rev Pharmacol Toxicol.201252: 381-400), and also can inhibit the growth of tumor cells by promoting the viability of p53, causing growth arrest and apoptosis of tumor cells (Jones RG et al, Mol cell 200518 (3): 283-93). Therefore, AMPK activators have an important role in the prevention and treatment of tumors.

Besides tumors, AMPK is also closely related to diabetes. It has been found that AMPK activity is significantly inhibited in peripheral tissues of obese mice and type II diabetic patients (violet b.et al, Crit Rev Biochem Mol biol 201045 (4): 276-95). Activation of AMPK promotes transfer of GLUT4, a glucose transporter in muscle, to the Cell membrane, and increases glucose absorption and catabolism in blood by muscle, thereby lowering blood glucose (Huang S, Czech MP. Cell Metab.20075 (4): 237-52). In the liver, AMPK can also promote the latter's enucleation by means of phosphorylation of CRTC2 or FOXO1 by means of phosphorylation of deacetylase HDAC4/5/7, both of which result in inhibition of the hepatic gluconeogenesis pathway and reduction of blood glucose (Altarejos, MontminM. Nat Rev Mol Cell biol. 201112 (3): 141-51). Meanwhile, the activation of AMPK can also promote fat hydrolysis and fatty acid oxidation of obese mice, thereby achieving the effects of reducing the fat content of the liver, treating fatty liver, or reducing the volume of fat tissues and losing weight (Garcia D et al, Cell Rep.201926 (1): 192-349; Pollard A et al, Nature Metab.20191: 340-349). Therefore, AMPK activators play an important role in the preparation of drugs for reducing fatty acid synthesis, drugs for inhibiting the synthesis of triglycerides and cholesterol, and drugs for preventing and/or treating obesity and type II diabetes.

Meanwhile, since AMPK has a multifunctional role in carbohydrate, fat and cholesterol metabolism and biosynthesis, which are closely related to parkinson's disease and alzheimer's disease (nat. rev. mol. Cell biol.2014,15, 634. 646), and prolonging the life of organisms (curr. biol.2007,17, 1646. cndot. 1656, Cell metab.2013,17, 101. cndot. 112, Cell metab.201420, 10-25, and nat. commun.2013,4,2192), etc., AMPK activators play an important role in the preparation of drugs for preventing and/or treating parkinson's disease, drugs for preventing and/or treating alzheimer's disease, or drugs for prolonging the life of mammals.

However, despite such important associations of AMPK with metabolic regulation and even human health, few AMPK activators are available clinically, and only metformin is currently used clinically as a first-line drug for type II diabetes. The target organs for metformin action are quite limited: can only act on the liver and the kidney; while AMPK has no regulating effect on tissues such as fat and muscle, which are closely related to the metabolic regulation effect. Therefore, the search for a wider range of AMPK activators has been a hot problem in academia and industry, and it is urgently needed to design and search for AMPK activators having novel structure, high safety and higher activity through a new idea.

Disclosure of Invention

The inventor of the invention designs and synthesizes a series of polysubstituted fluorine-containing imidazole salt derivatives with novel structure, high safety and high activity through extensive and intensive research in order to search for a new AMPK activator, and researches the influence of the novel derivatives on an AMPK signal path.

The present invention provides compounds of the general formula:

or a stereoisomer of the above compound, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

Wherein the definitions of the substituents and symbols are explained in detail below.

An object of the present invention is to provide a class of compounds having an activity of activating 5' -adenosine monophosphate-activated protein kinase (AMPK), and stereoisomers, prodrugs, pharmaceutically acceptable salts, or pharmaceutically acceptable solvates thereof.

Another object of the present invention is to provide a process for the preparation of the above compounds.

It is another object of the present invention to provide a pharmaceutical composition comprising the above compound.

Another object of the present invention is to provide the use of the above-mentioned compound and a pharmaceutical composition comprising the same for the preparation of a medicament for activating 5' -adenosine monophosphate-activated protein kinase (AMPK) activity.

Another object of the present invention is to provide the use of the above-mentioned compound and a pharmaceutical composition comprising the same for the preparation of a medicament for reducing fatty acid synthesis, a medicament for inhibiting triglyceride and cholesterol synthesis, a medicament for preventing and/or treating obesity and type II diabetes, a medicament for preventing and/or treating tumors, a medicament for preventing and/or treating parkinson's disease, a medicament for preventing and/or treating alzheimer's disease, or a medicament for prolonging the lifespan of a mammal.

Drawings

Figure 1 is a graph of AMPK activation levels in MEF cells by representative compounds. FIG. 1 illustrates that compounds activate AMPK in Mouse Embryonic Fibroblasts (MEFs). The results show that the tested compound can effectively activate AMPK at 10nM, promote phosphorylation of AMPK (p-AMPK) and phosphorylation of ACC1/ACC2 (p-ACC) which is a downstream substrate of AMPK.

FIG. 2 illustrates the possible metabolic pathways of LXY-Cl in human hepatocyte incubation systems. The results show that LXY-Cl can be metabolized into a plurality of different metabolites in human liver cells, and the residual relative content of the original drug is 21% after incubation for 120 minutes.

FIG. 3 illustrates the possible metabolic pathways of IB-33 in a human hepatocyte incubation system. The results show that IB-33, i.e. the fluorine-containing substituted compound corresponding to LXY-Cl, is relatively stable in human liver cells, and the residual relative content of the original drug is 79% after incubation for 120 minutes. Comparison of the metabolite data of IB-33 and LXY-Cl in hepatocytes shows a significant increase in the metabolic stability of the fluorine-containing compounds.

Detailed Description

Various specific embodiments, modes and examples are described herein, including exemplary embodiments and definitions employed for understanding the claimed invention. While the following detailed description gives specific preferred embodiments, those skilled in the art will appreciate that these embodiments are merely exemplary, and that the present invention can be practiced in other ways. For infringement purposes, the scope of the invention will refer to any one or more of the appended claims, including equivalents thereof, as well as elements or limitations that are equivalent to those that are recited.

The invention is realized by the following technical scheme.

In a first aspect, the present invention provides a compound of the general formula:

or a stereoisomer of the above compound, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof,

wherein the content of the first and second substances,

R₁selected from C10-C20 alkyl substituted with 1-15 fluorine atoms;

R₂selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl;

R₃is selected from

1)

Wherein Z₁，Z₂，Z₃，Z₄，Z₅Each independently selected from:

(1) hydrogen, fluorine, chlorine, bromine, iodine, nitro, cyano, amino, hydroxyl, hydroxycarbonyl, methoxyformyl, ethoxyformyl, N-propoxycarbonyl, isopropoxyformyl, carbamoyl, N-methylcarbonyl, N-ethylformyl, N-N-propylformyl, N-isopropylformyl, N-cyclopropylformyl, N-N-butylformyl, N-isobutylformyl, N-tert-butylformyl, N-cyclobutylformyl, N-N-pentylformyl, N-isopentylcarbonyl, N-cyclopentylcarbonyl, N-N-hexylformyl, N-isohexylcarbonyl, N-cyclohexylformyl, N, N-dimethylformyl, N, N-diethylformyl, n, N-di-N-propylformyl, N, N-diisopropylformyl, cyclopropylcarbamoyl, cyclobutylaminoformyl, cyclopentylaminoformyl, cyclohexylcarbamoyl, 4-hydroxypiperidinoylformyl, piperazinoylformyl, 4-N-methylpiperazinoylformyl, 4-N-ethylpiperazinoylformyl, 4-N-N-propylpiperazinoylformyl, 4-N-isopropylpiperazinoylformyl, methylsulfonyl, ethylsulfonyl, N-propylsulfonyl, isopropylsulfonyl, N-butylsulfonyl, isobutylsulfonyl, hydroxysulfonyl, aminosulfonyl, N-methylsulfonyl, N-ethylsulfonyl, N-N-propylsulfonyl, N-isopropylsulfonyl, N-cyclopropylsulfonyl, N-N-butylsulfonyl group, N-isobutylsulfonyl group, N-tert-butylsulfonyl group, N-cyclobutylsulfonyl group, N-N-pentylsulfonyl group, N-isopentylsulfonyl group, N-cyclopentylsulfonyl group, N-N-hexylsulfonyl group, N-isohexylsulfonyl group, N-cyclohexylsulfonyl group, N, N-dimethylsulfonyl group, N, N-diethylsulfonyl group, N, N-di-N-propylsulfonyl group, N, N-diisopropylsulfonyl group, cyclopropylaminosulfonyl group, cyclobutylaminosulfonyl group, cyclopentaminylsulfonyl group, cyclohexylaminosulfonyl group, 4-hydroxypiperidinylsulfonyl group, piperazinylsulfonyl group, 4-N-methylpiperazinylsulfonyl group, 4-N-ethylpiperazinylsulfonyl group, 4-N-N-propylpiperazinylsulfonyl group, 4-N-isopropylpiperazinylsulfonyl, carboxamido, acetamido, propionamido, N-butylamido, isobutyramido, cyclopropylcarboxamido, cyclobutylcarboxamido, cyclopentylcarboxamido, cyclohexylcarboxamido, methanesulfonamido, ethanesulfonamide, N-propanesulfonamide, isopropylsulfonamido, N-butylsulfonamido, isobutylsulfonamido;

(2) C1-C6 alkyl, C1-C6 alkoxy, C1-C6 oxygen-containing alkyl, C1-C6 fluorine-containing alkyl, C1-C6 fluorine-containing alkoxy;

(3)Z₂and Z₃May form an oxygen-containing substituted or unsubstituted five-or six-membered ring; the substituents may be selected from the group consisting of₁The same substituents;

(4)Z₄and Z₅A nitrogen-containing substituted or unsubstituted five-or six-membered ring may be formed; the substituents may be selected from the group consisting of₁The same substituents;

Z₆selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl;

2)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

3)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

4)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

X^-is anion of inorganic acid salt or organic acid salt which can be accepted in pharmacy.

In some embodiments, R₁Selected from: C14-C18 alkyl substituted by 1,3, 5, 7, 9, 11, 13 or 15 fluorine atoms.

In some embodiments, R₁Selected from: c₁₆FH₃₂-、C₁₄F₇H₂₂-、C₁₅F₉H₂₂-、C₁₆F₁₁H₂₂-、C₁₇F₁₃H₂₂-。

In some embodiments, R₂Selected from: hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl.

In some embodiments, R₂Selected from: hydrogen, methyl, isopropyl, cyclopropyl.

In some embodiments, R₃Is composed of

Wherein Z₁，Z₂，Z₃，Z₄，Z₅Each of 2 of which is independently selected from the following, the remainder being hydrogen:

(1) hydrogen, fluorine, chlorine, bromine, iodine, nitro, cyano, amino, hydroxy, hydroxycarbonyl, carbamoyl, methanesulfonyl, hydroxysulfonyl, aminosulfonyl, carboxamido, methanesulfonamide;

Z₆selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl; preferably Z₆Is hydrogen or methyl.

In some embodiments, R₃Is composed of

Wherein Z₁，Z₂，Z₄，Z₅2 of (a) are each independently selected from the group consisting of₃Is hydrogen:

In some embodiments, R₃Is composed of

Wherein Z is₁，Z₂，Z₃，Z₄，Z₅Z in (1)₁，Z₅Each, or Z₂，Z₄Each, or Z₁，Z₄Each independently selected from the following, the remainder being hydrogen:

In some embodiments, X^-Is chloride ion, bromide ion, iodide ion, sulfate ion, phosphate ion, maleate ion, fumarate ion, tartrate ion, palmitate ion, oxalate ion, citrate ion, succinate ion, methanesulfonate ion, benzenesulfonate ion, p-toluenesulfonate ion.

In a second aspect, the present invention provides the following compounds:

or a stereoisomer thereof, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof

Wherein R is₁Selected from C10-C20 alkyl substituted with 1-15 fluorine atoms;

R₂selected from: hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl;

S₁selected from: 1 ' -halo, 1 ' -C1-C6 alkoxy (preferably 1 ' -C1-C3 alkoxy);

S₂selected from: 4 '-halo, 5' -halo;

X^-is pharmaceutically acceptable inorganic acid and organic acidAn anion.

In some embodiments, R₁Selected from: n-C₁₆FH₃₂-、n-C₁₄F₇H₂₂-、n-C₁₅F₉H₂₂-、n-C₁₆F₁₁H₂₂-、 n-C₁₇F₁₃H₂₂-。

In some embodiments, R is preferred₂Selected from: hydrogen, C1-C3 alkyl, C3-C4 cycloalkyl.

In some embodiments, S₁,S₂Each is as follows: 1 ' -halogen, 5 ' -halogen, or 1 ' -C1-C6 alkoxy (preferably 1 ' -C1-C3 alkoxy), 4 ' -halogen.

In a third aspect, the present invention provides the following compounds:

wherein, for R₁、R₂、R₃And X^－Wherein n is 9 to 19,

Unless otherwise specified, the above groups and substituents have the ordinary meaning in the field of pharmaceutical chemistry.

“C₁₀-C₂₀Alkyl "includes straight or branched chain groups having any two integers between 10 and 20 carbon atoms as endpoints. For example, "C₁₀-C₂₀Alkyl "includes C14-C18 alkyl, C₁₀-C₁₈Alkyl radical, C₁₀-C₁₆Alkyl radical, C₁-C₄Alkyl radical, C₂-C₂₀Alkyl radical, C₂-C₁₆Alkyl radical, C₆-C₂₀Alkyl radical, C₆-C₁₆Alkyl, and the like, listed above by way of example only, and not by way of limitation with respect to the stated interval.

The term "C10-C20 alkyl substituted by 1 to 15 fluorine atoms" means the above-mentioned "C10-C20 alkyl group₁₀-C₂₀Alkyl "is substituted with 1, 2, 3,4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 fluorine atoms.

The term "C₁-C₆Alkyl "refers to any straight or branched chain group containing 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, tert-pentyl, n-hexyl, and the like.

The term "C₁-C₄Alkyl "refers to any straight or branched chain group containing 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and the like.

The term "C₁-C₃Alkyl "refers to any straight or branched chain group containing 1 to 3 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, and the like.

The term "C₃-C₆Cycloalkyl "refers to a 3-to 6-membered all-carbon monocyclic ring, which may contain 0, one or more double bonds, but does not have a fully conjugated pi-electron system. Examples of cycloalkyl radicalsExamples are, but not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene.

The term "C3-C4 cycloalkyl" refers to a 3-to 4-membered all-carbon monocyclic ring, examples of which are, but are not limited to, cyclopropane, cyclobutane.

The term "halogen" refers to fluorine, chlorine, bromine, iodine.

The term "cyano" refers to the residue — CN.

The term "nitro" refers to-NO₂A group.

The terms "alkoxy", "cyclyloxy" and derivatives thereof refer to any of the above alkyl groups (e.g., C)₁-C₂₄Alkyl radical, C₁-C₆Alkyl, etc.), cycloalkyl (e.g. C)₃-C₆Cycloalkyl) which is linked to the rest of the molecule through an oxygen atom (-O-).

From all the above descriptions, it is obvious to the person skilled in the art that any group whose name is a compound name, such as "fluorine-containing oxyalkyl", shall mean that the moiety conventionally derived therefrom is constructed, for example, from oxyalkyl substituted by fluorine group, wherein alkyl is as defined above.

The term "oxygen-containing substituted or unsubstituted five-or six-membered ring" or "nitrogen-containing substituted or unsubstituted five-or six-membered ring" refers to a 5-or 6-membered saturated or partially unsaturated carbocyclic ring in which one or more carbon atoms are replaced by oxygen or nitrogen. Non-limiting examples are, for example, pyrane, pyrrolidine, pyrroline, imidazoline, imidazolidine, pyrazolidine, pyrazoline, dihydrofuran, tetrahydrofuran, 1, 3-dioxolane, piperidine, piperazine, morpholine, tetrahydropyrrolyl, and the like.

In the above for R₃Z in (1)₁，Z₂，Z₃，Z₄，Z₅In the definition of "wherein, Z₁，Z₂，Z₃，Z₄， Z₅Z in (1)₁，Z₅Each, or Z₂，Z₄Each, or Z₁，Z₄Each independently selected from the following, the remainder being hydrogen: (1) hydrogen, fluorineChlorine, bromine, iodine, nitro, cyano; (2) C1-C3 alkyl, C1-C6 alkoxy, C1-C6 oxyalkyl, C1-C6 fluoroalkyl, C1-C6 fluoroalkoxy ", which means: "Z" is₁，Z₅Each independently selected from the group consisting of₁， Z₅Independently are each independently exemplified by "(1) hydrogen, fluoro, chloro, bromo, iodo, nitro, cyano; (2) any combination of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 oxygen-containing alkyl, C1-C6 fluorine-containing alkyl and C1-C6 fluorine-containing alkoxy, and' Z₂，Z₄Each independently selected from the group consisting of₂，Z₄Independently are each independently exemplified by "(1) hydrogen, fluoro, chloro, bromo, iodo, nitro, cyano; (2) any combination of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 oxygen-containing alkyl, C1-C6 fluorine-containing alkyl and C1-C6 fluorine-containing alkoxy, and' Z₁，Z₄Each independently selected from the group consisting of₁，Z₄Independently are each independently exemplified by "(1) hydrogen, fluoro, chloro, bromo, iodo, nitro, cyano; (2) any combination of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 oxygen-containing alkyl, C1-C6 fluorine-containing alkyl and C1-C6 fluorine-containing alkoxy.

As used herein, unless otherwise specified, the term "prodrug" refers to a derivative that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide a compound of the invention. Prodrugs undergo this reaction only under biological conditions to become active compounds, or they are active in a form in which they do not react. Prodrugs can generally be prepared using well known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery (1995) 172-.

As used herein, examples of the term "pharmaceutically acceptable salts of compounds of formula (I)" are organic acid addition salts formed from organic acids that form pharmaceutically acceptable anions, including but not limited to formate, acetate, propionate, benzoate, maleate, fumarate, succinate, tartrate, citrate, ascorbate, α -ketoglutarate, α -glycerophosphate, alkylsulfonate or arylsulfonate; preferably, the alkyl sulfonate is a methyl sulfonate or an ethyl sulfonate; the aryl sulfonate is benzene sulfonate or p-toluene sulfonate. Suitable inorganic salts may also be formed and include, but are not limited to, hydrochlorides, hydrobromides, hydroiodides, nitrates, bicarbonates and carbonates, sulfates or phosphates, and the like.

Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficient amount of a basic compound with a suitable acid to provide a pharmaceutically acceptable anion.

The term "treating" as used herein generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of preventing the disease or its symptoms, in whole or in part; and/or may be therapeutic in terms of partially or completely stabilizing or curing the disease and/or side effects due to the disease. As used herein, "treatment" encompasses any treatment of a disease in a patient, including: (a) preventing a disease or condition in a patient susceptible to the disease or condition but not yet diagnosed as having the disease; (b) inhibiting the symptoms of the disease, i.e., arresting its development; or (c) alleviating the symptoms of the disease, i.e., causing regression of the disease or symptoms.

According to a specific embodiment of the present invention, the compound, a stereoisomer thereof, a prodrug thereof, or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, wherein the compound is one of the compounds described in the following examples.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound according to any one of the above technical schemes, a stereoisomer thereof, a prodrug thereof, or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

Methods of preparing various pharmaceutical compositions containing certain amounts of active ingredients are known or will be apparent to those skilled in the art in view of this disclosure. The process for preparing the PHARMACEUTICAL composition comprises incorporating suitable PHARMACEUTICAL excipients, carriers, diluents and the like, as described in REMINGTON' S PHARMACEUTICAL SCIENCES, Martin, e.w., ed., Mack Publishing Company,19th ed. (1995).

The pharmaceutical formulations of the present invention are manufactured in a known manner, including conventional mixing, dissolving or lyophilizing processes. The compounds of the invention may be formulated into pharmaceutical compositions and administered to a patient by a variety of routes appropriate for the mode of administration selected, for example, orally or parenterally (by intravenous, intramuscular, topical or subcutaneous routes).

Thus, the compounds of the present invention may be administered systemically, e.g., orally, in combination with a pharmaceutically acceptable carrier, such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules and may be compressed into tablets. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of swallowable tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The proportions of such compositions and formulations can, of course, vary and can range from about 1% to about 99% by weight of a given unit dosage form. In such therapeutically useful compositions, the amount of active compound is such that an effective dosage level is obtained.

Tablets, troches, pills, capsules and the like may also contain: binders, such as gum tragacanth, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; disintegrating agents, such as corn starch, potato starch, alginic acid, and the like; lubricants, such as magnesium stearate; and sweeteners such as sucrose, fructose, lactose or aspartame; or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavor. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a vegetable oil or polyethylene glycol. Various other materials may be present, as coatings, or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules may be coated with gelatin, wax, shellac, or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl or propylparabens as preservatives, a dye and a flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compounds may be incorporated into sustained release formulations and devices.

The active compounds may also be administered intravenously or intraperitoneally by infusion or injection. An aqueous solution of the active compound or its salt may be prepared, optionally mixed with a non-toxic surfactant. Dispersions in glycerol, liquid polyethylene glycols, glycerol triacetate and mixtures thereof, and oils may also be prepared. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders of the active ingredient, optionally encapsulated in liposomes, containing ready-to-use preparations of injectable or infusible solutions or dispersions suitable for sterility. In all cases, the final dosage form must be sterile, liquid and stable under the conditions of manufacture and storage. The liquid carrier can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), vegetable oil, non-toxic glycerol esters, and suitable mixtures thereof. Suitable fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersants, or by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use of compositions which delay absorption of the agent (e.g., aluminum monostearate and gelatin).

Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional required ingredients previously present in the sterile-filtered solution.

Useful solid carriers include finely divided solids (e.g., talc, clay, microcrystalline cellulose, silicon dioxide, alumina, and the like). Useful liquid carriers include water, ethanol or ethylene glycol or water-ethanol/ethylene glycol mixtures in which the compounds of the present invention may be dissolved or dispersed in effective amounts, optionally with the aid of non-toxic surfactants. Adjuvants (such as fragrances) and additional antimicrobial agents may be added to optimize the properties for a given use.

Thickeners (e.g., synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified inorganic materials) can also be used with liquid carriers to form coatable pastes, gels, ointments, soaps, etc., for direct application to the skin of the user.

The therapeutically required amount of the compound or its active salt or derivative will depend not only on the particular salt selected, but also on the mode of administration, the nature of the disease to be treated and the age and condition of the patient, and will ultimately be at the discretion of the attendant physician or clinician.

The formulations may be presented in unit dosage form comprising physically discrete units of a unit dose suitable for administration to the human or other mammalian body. The unit dosage form may be a capsule or tablet, or a plurality of capsules or tablets. The amount of unit dose of the active ingredient may vary or be adjusted from about 0.1 to about 1000 mg or more, depending upon the particular treatment involved.

In addition, the application of various new pharmaceutical forms such as cream masses, microspheres and nanospheres is also included, such as those prepared using microparticulate dispersion systems including polymeric micelles (polymeric micelles), nanoemulsions (nanoemulsions), submicroemulsions (microcapsules), microspheres (microspheres), liposomes (lipomes), and niosomes (also known as nonionic surfactant vesicles).

In another aspect, the present invention further provides a method for preparing the compound according to any one of the above technical schemes, comprising the following steps:

route one:

reaction conditions are as follows: (a) substitution reaction of brominated hydrocarbons; (b) substitution of brominated hydrocarbons.

And a second route:

reaction conditions are as follows: (a) substitution reaction of brominated hydrocarbons under alkaline conditions (such as sodium hydride, sodium tert-butyl alkoxide, etc.); (b) substitution of brominated hydrocarbons.

In another aspect, the present invention also provides a use of the compound, the stereoisomer, the prodrug, the pharmaceutically acceptable salt or the pharmaceutically acceptable solvate thereof according to any one of the above technical schemes, and a pharmaceutical composition containing the compound in preparing a medicament for inhibiting cholesterol synthesis, a medicament for reducing fatty acid synthesis, a medicament for preventing and/or treating diabetes, a medicament for preventing and/or treating tumor, a medicament for preventing and/or treating parkinson's disease, a medicament for preventing and/or treating alzheimer's disease, or a medicament for prolonging the lifespan of a mammal.

Experimental part

For the examples referred to below, the compounds of the invention are synthesized using the methods described herein or other methods well known in the art.

Universal purification and analysis methods

Thin layer chromatography was performed on silica gel GF254 pre-coated plates (Qingdao oceanic chemical plant). Column chromatography over silica gel (300-400 mesh, Shintai yellow silica gel development reagent factory) at medium pressure or by using an ISCO Combiflash Rf200 Rapid purification System with pre-loaded silica gel cartridges (ISCO or Welch). The components were developed by UV light (lambda: 254nm) and by iodine vapor. When necessary, the compounds were purified by preparative HPLC preparation on a Waters Symmetry C18 (19X 50mm,5 μm) column or on a Waters X Terra RP18 (30X 150mm,5 μm) column using a Waters preparative HPLC 600 equipped with a 996 Waters PDA detector and Micromass mod. The method comprises the following steps: phase A0.1% TFA/MeOH 95/5; phase B MeOH/H2O 95/5. Gradient: performing the treatment for 10-90% B for 8min, and maintaining for 90% B for 2 min; the flow rate was 20 mL/min. The method 2 comprises the following steps: phase A0.05% NH4OH/MeOH 95/5; phase B MeOH/H2O 95/5. Gradient: 10-100% B for 8min, and keeping 100% B for 2 min. The flow rate was 20 mL/min.

Will be provided with¹H-NMR spectra were recorded in DMSO-d6 or CDCl3 on a Bruker Avance 600 spectrometer (for 1H) operating at 600 MHz. The residual solvent signal was used as a reference (═ 2.50 or 7.27 ppm). Chemical shifts () are reported in parts per million (ppm) and coupling constants (J) are in Hz. The following abbreviations are used for peak splitting, s is mono; br.s. ═ wide signal; d is bis; t is three; m is multiple; dd is bis-bis.

Electrospray (ESI) mass spectra were obtained via Finnigan LCQ ion trap.

Unless otherwise indicated, all final compounds were homogeneous (not less than 95% pure) as determined by High Performance Liquid Chromatography (HPLC). HPLC-UV-MS analysis for the evaluation of compound purity was performed by combining an ion trap MS apparatus with an HPLC system SSP4000(Thermo Separation Products) equipped with an autosampler LC Pal (CTC analytical) and a UV6000LP diode array detector (UV detection 215-400 nm). The device control, data acquisition and processing was performed with xcalibur1.2 software (Finnigan). HPLC chromatography was performed at room temperature and 1mL/min flow rate using a Waters X Terra RP18 column (4.6X50 mm; 3.5 μm). Mobile phase a is ammonium acetate 5mM buffer (pH 5.5 with acetic acid) acetonitrile 90:10, mobile phase B ammonium acetate 5mM buffer (pH 5.5 with acetic acid) acetonitrile 10: 90; the gradient was run from 0 to 100% B for 7 minutes, then 100% B was held for 2 minutes before re-equilibration.

Reagent Purification is described in the paper of Purification of Laboratory Chemicals (Perrin, D.D., Armarego, W.L.F. and Perrin Eds, D.R.; Pergamon Press: Oxford, 1980). The petroleum ether is 60-90 deg.C fraction, and the ethyl acetate, methanol and dichloromethane are analytically pure.

Detailed Description

The embodiments of the present invention are described in detail below by way of specific examples, but they should not be construed as limiting the invention in any way.

The compounds of the general formula are prepared by two kinds of synthesis.

Wherein the content of the first and second substances,

synthetic route of compound IA

Weighing diethylaminosulfur trifluoride DAST (2eq) in a reaction bottle, dissolving a compound A (1eq) in dichloromethane, adding the dichloromethane at normal temperature into the reaction bottle, sealing, pumping nitrogen, placing the mixture in a 40 ℃ oil bath, stirring for 8 hours, cooling a reaction system to room temperature, pouring the mixture into ice water, extracting the ice water with dichloromethane, and sequentially using saturated NaHCO for an organic phase₃The solution was washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered, the organic phase was concentrated and chromatographed on silica gel (dichloromethane elution) to obtain compound B.

Dissolving the compound B (1eq) and the compound C (1eq) in THF, adding sodium tert-butoxide (2eq) under stirring in an ice bath, continuously stirring for 20min, placing in an oil bath at 40 ℃, stirring, cooling the system to room temperature after TLC monitoring reaction is completed, adding a saturated ammonium chloride solution and ethyl acetate for extraction, washing an organic phase once with water and saturated sodium chloride, drying with anhydrous sodium sulfate, filtering, concentrating the organic phase, and performing silica gel column chromatography (dichloromethane/methanol) to obtain the compound D.

Mixing Compounds D (1eq) and R₃-X (1.2eq) in acetonitrileAdding potassium iodide (6 eq) which may or may not be optionally present, sealing, placing in a 70 ℃ oil bath, stirring for 12h, cooling the system to room temperature, concentrating, and performing silica gel column chromatography (dichloromethane/methanol) to obtain compound IA.

The specific implementation of scheme one of the synthesis of compound I is described below.

1. Compound IA-1:

dissolving 16-bromo-1-hexadecanol (1606mg,5mmol) (CAS:59101-28-9, Jiangsu Aikang, Jiangsu) in 4 mL of dichloromethane, dropwise adding into a microwave tube filled with diethylaminosulfur trifluoride (1612mg, 10mmol) (CAS:38078-09-0, Annaiji, Shanghai), sealing, introducing nitrogen, stirring at 40 ℃ in an oil bath for 8h, cooling the reaction system to room temperature, pouring into ice water, extracting with dichloromethane, and sequentially using saturated NaHCO as an organic phase₃Washing the solution and a saturated sodium chloride solution once, drying the solution by using anhydrous sodium sulfate, filtering, concentrating an organic phase, performing silica gel column chromatography (eluting by using dichloromethane), directly dissolving an obtained product and 2-methylimidazole (410mg and 5mmol) in a raw material A (attached table 1) in 30mL THF, adding sodium tert-butoxide (960mg and 10mmol) under ice-bath stirring (CAS:865-48-5, Annaiji, Shanghai), continuously stirring for 20min, placing the mixture in an oil bath at 40 ℃ for stirring, cooling a system to room temperature after TLC monitoring reaction is completed, adding a saturated ammonium chloride solution and ethyl acetate for extraction, washing the organic phase once by using water and saturated sodium chloride, drying the anhydrous sodium sulfate, filtering, concentrating the organic phase, and performing silica gel column chromatography (dichloromethane/methanol) to obtain an intermediate int-1(106mg and 0.326 mmol).

The intermediate int-1(35.4mg,0.11mmol) and 2, 6-dichlorobenzyl chloride (22.5mg, 0.12mmol) in the raw material B (attached table 1) were dissolved in acetonitrile, and after sealing, the mixture was stirred in a 70 ℃ oil bath for 12 hours, and the system was cooled to room temperature, concentrated, and subjected to silica gel column chromatography (dichloromethane/methanol) to obtain compound IA-1(21.6 mg).

2. Compounds IA-2 to IA-3 were synthesized using a similar procedure and the corresponding starting materials are shown in attached Table 1.

Synthetic route of compound I

Placing compound A (1eq), compound B (1.2eq) and AIBN (0.1eq) in a microwave tube, sealing, pumping nitrogen for 3 times, placing in an oil bath at 60 ℃ or 100 ℃ for reaction for 24 hours, cooling to room temperature, adding glacial acetic acid and zinc powder (20eq) into the system, stirring at room temperature for 12 hours, stopping the reaction, filtering the system, concentrating, pouring proper amount of water, adjusting the pH to be neutral by using 2N sodium hydroxide solution, extracting the water phase by using petroleum ether, and concentrating the organic phase to obtain compound C.

Dissolving a compound C (1eq) and triethylamine (3eq) in dichloromethane, placing the solution in an ice bath for stirring, slowly and dropwise adding methylsulfonyl chloride (2eq), continuing to stir in the ice bath for 10min after the feeding is finished, placing the solution at normal temperature for reacting for 10h, adding water for quenching, extracting with dichloromethane/water, washing an organic phase with water and saturated sodium chloride in sequence, drying with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography (petroleum ether/ethyl acetate) to obtain a compound D.

And dissolving the compound D (1eq) and lithium bromide (3eq) in acetone, placing the system at 60 ℃ for reflux reaction for 12h, filtering, concentrating, and performing silica gel column chromatography (petroleum ether elution) to obtain a compound E.

Dissolving compounds F (1eq) and E (1.2eq) in THF, adding sodium tert-butoxide (2eq) under stirring in an ice bath, continuously stirring for 20min, placing in an oil bath at 40 ℃, stirring, cooling the system to room temperature after TLC monitoring reaction is completed, adding a saturated ammonium chloride solution and ethyl acetate for extraction, washing an organic phase once with water and saturated sodium chloride, drying with anhydrous sodium sulfate, filtering, concentrating the organic phase, and performing silica gel column chromatography (dichloromethane/methanol) to obtain a compound G.

Mixing the compounds G (1eq) and R₃-X (1.2eq) is dissolved in acetonitrile, potassium iodide (6 eq) optionally present or absent is added, after sealing, stirred in a 70 ℃ oil bath for 12h, the system is cooled to room temperature, concentrated, and chromatographed on silica gel (dichloromethane/methanol) to obtain compound IB.

The implementation of scheme one of the synthesis of compound IB is described below.

1. Compound IB-1:

10-undecen-1-ol (3406mg, 20mmol) (CAS:112-43-6, Jiangsu Aikang, Jiangsu), perfluoroiodopropane (7102mg, 24mmol) and AIBN (328.4mg, 2mmol) (CAS: 78-67-1, exploration, Shanghai) in raw material C (attached Table 1) were placed in a microwave tube, sealed, nitrogen was pumped out 3 times, placed in an oil bath at 60 ℃ for 24 hours, cooled to room temperature, 70ml of glacial acetic acid and zinc powder (26g, 400mmol) (CAS:7440-66-6, Lvjinghi, Fujiangmen) were added to the system, stirred at room temperature for 12 hours, the reaction was stopped, filtered, concentrated, poured in appropriate amount of water, adjusted to neutral pH with 2N sodium hydroxide solution, the aqueous phase was extracted with petroleum ether, the organic phase was concentrated, and intermediate int-2(3820mg, 11.25mmol) was obtained.

Int-2(3820mg, 11.25mmol) and triethylamine (3408mg, 33.75mmol) (CAS:121-44-8, Annaiji, Shanghai) are dissolved in 100mL of dichloromethane, the solution is placed in an ice bath and stirred, methanesulfonyl chloride (7735 mg, 67.5mmol) (CAS:124-63-0, Annaiji, Shanghai) is slowly added dropwise, after the feeding is finished, the ice bath is continued and stirred for 10min, the mixture is placed at normal temperature for reaction for 10h, water is added for quenching, dichloromethane/water extraction is carried out, an organic phase is washed once by water and saturated sodium chloride, anhydrous sodium sulfate is dried and then concentrated, and silica gel column chromatography (petroleum ether/ethyl acetate) is carried out to obtain an intermediate int-3(4354mg, 10.4 mmol).

Int-3(4354mg, 10.4mmol) and lithium bromide (2709mg, 31.2mmol) (CAS:7550-35-8, Annaiji, Shanghai) were dissolved in 50mL of acetone, the system was placed at 60 ℃ for reflux reaction for 12h, filtered, concentrated, and chromatographed on a silica gel column (petroleum ether elution) to give intermediate int-4(3715mg, 9.2 mmol).

Int-4(403.2mg, 1mmol) and imidazole (81.7mg, 1.2mmol) in raw material A (attached Table 1) are dissolved in 30mL THF, sodium tert-butoxide (192mg, 2mmol) is added under ice-bath stirring, the mixture is placed in an oil bath at 40 ℃ after stirring for 20min, the system is cooled to room temperature after TLC monitoring reaction is completed, saturated ammonium chloride solution and ethyl acetate are added for extraction, the organic phase is washed once with water and saturated sodium chloride, anhydrous sodium sulfate is dried and then filtered, the organic phase is concentrated, and silica gel column chromatography (dichloromethane/methanol) is carried out to obtain an intermediate int-5(295mg, 0.75 mmol).

Int-5(39mg, 0.1mmol) and 2, 6-dichlorobenzyl chloride (23.4mg, 0.12mmol) in raw material B (attached Table 1) were dissolved in 1.5mL of acetonitrile, and after sealing, the mixture was stirred in a 70 ℃ oil bath for 12 hours, and the system was cooled to room temperature, concentrated, and subjected to silica gel column chromatography (dichloromethane/methanol) to obtain compound IB-1(20.5 mg).

2. Other compounds IB-1 to IB-58 can be synthesized using similar methods, and the corresponding starting materials are shown in attached Table 1.

TABLE 1 attached partial commercial raw materials for the synthesis of the example compounds

Starting materials A
	Imidazole, CAS 288-32-4, Meclin, Shanghai
2-methylimidazole, CAS:693-98-1, Acros Organics, Belgium
	2-Ethylimidazole, CAS:1072-62-4, Meclin, Shanghai
2-isopropylimidazole, CAS:36947-68-9, fructus Piperis, Shanghai
	2-tert-butylimidazole, CAS:36947-69-0, Shaoyuan, Shanghai
2-cyclopropyl-1H-imidazole, CAS 89532-38-7, Wash-Wikipedia, Beijing

Raw material B
	2-chloro-6-fluorobenzyl bromide CAS 68220-26-8, Diploc, Shanghai
2-chloro-6-fluorobenzyl chloride, CAS:55117-15-2, Annaiji, Shanghai
	2, 6-dichlorobenzyl bromide, CAS 20443-98-5, Annaiji, Shanghai
2, 6-dichlorobenzyl chloride, CAS:2014-83-7, Annaiji, Shanghai
	5-fluoro-2-methoxybenzyl bromide, CAS:20-3-560364, Annaiji, Shanghai

	Raw material C
Perfluoro iodopropane, CAS:754-34-7, Jiangsu Aikang, Jiangsu
	Perfluoro iodobutane, CAS:423-39-2, Jiangsu Aikang, Jiangsu
Perfluoro iodopentane, CAS 638-79-9, Jiangsu Aikang and Jiangsu
	Perfluoro hexyl iodoalkaneCAS 355-43-1, Annaiji, Shanghai

TABLE 1 Structure and characterization of Compounds IA-IB

Test examples

And (3) biological activity test:

1. cellular level AMPK activity assay

The test of the ability of the compound to activate AMPK in Mouse Embryo Fibroblasts (MEFs) can be realized by detecting the phosphorylation level of threonine at position 172 of AMPK (p-AMPK alpha) and the phosphorylation level of serine at position 79 of ACC1/ACC2 (p-ACC) which is a substrate of AMPK by an immunoblotting (western blot) method (figure 1 and table I).

The specific method comprises the following steps:

(1) MEFs with loxP insert or wild type were plated in six-well plates and cultured in DMEM containing 10% serum. If a certain gene needs to be knocked out, adenovirus capable of expressing cre is added into the culture hole when the density of corresponding MEFs with loxP insertion sequences reaches about 30%, and then the culture hole is cultured for more than 24 hours;

(2) when the cell density was close to 90%, the cells were replaced with fresh DMEM while adding the compound (final concentration 10nM) to the cells for 2 hours, with an equal volume of DMSO as a negative control and the cells treated with AICAR (3mM) added as a positive control;

(3) the culture medium was aspirated off, the cells were lysed with 200. mu.L of cell lysate (after formulation attachment), the cells were scraped from the culture dish, disrupted by sonication, and centrifuged at 20000g for 10 minutes at low temperature;

(4) mixing the supernatant with an equal volume of 2 × SDS solution (after formulation), running 8% SDS-PAGE, transferring the proteins to PVDF membranes, blocking each PVDF membrane with 25mL skim milk for 1 hour, and then rinsing with TBST buffer (after formulation) for 3 times, each for 10 minutes;

(5) AMPK α subunit primary antibodies (Cell Signaling Technology, #2532), AMPK threonine phosphorylation primary antibodies at position 172 (Cell Signaling Technology, #2535), ACC primary antibodies (Cell Signaling Technology, #3662), ACC serine phosphorylation primary antibodies at position 79 (Cell Signaling Technology, #3661) were expressed as follows: 1000 is diluted into a primary anti-dilution solution (after the formula is attached), reacts with a PVDF membrane at room temperature for 12 hours, and is rinsed for 3 times by using TBST buffer solution;

(6) adding 1: 1000 dilution of HRP-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch, 111-035-003), reaction at room temperature for 1 hour, and washing 3 times with TBST buffer;

(7) the PVDF membrane is wiped dry, reacts in ECL mixed solution (WesternBright ECL HRP substrate, Advansta), is exposed and developed by medical X-ray film, and is finally washed, dried and scanned to obtain the related data of AMPK activation.

The used reagent formula is as follows:

cell lysis solution: 20mM Tris-base, pH7.5, 150mM NaCl,1mM EDTA,1mM EGTA,2.5 mM Sodium pyrophosphate (Sodium pyrophosphate),1mM beta-glycerophosphate (beta-glycerophosphate), 1% Triton X-100 (v/v);

2 SDS solution: 20% Glycerol (Glycerol) (v/v), 4% SDS (m/v), 10% β -mercaptoethanol (β -mecaptoethanol) (v/v), 0.01% Bromophenol blue (Bromophenol blue) (m/v);

TBST buffer: 4.84% Tris-base (m/v), 8% NaCl (m/v), 0.1% Tween-20 (v/v);

primary anti-dilution solution: TBST buffer containing 5% BSA (v/v)

TABLE I ability of Compounds to activate AMPK in MEF cells

^aThe extent to which compounds activate AMPK in MEF cells is expressed as the fold of the ratio of p-AMPK α/AMPK α and p-ACC/ACC after treatment with compound (10nM) relative to positive control AICAR (3mM) (the corresponding brightness of the bands is quantified using Image J), see in particular fig. 1.

2. Liver microsome stability test

The experimental steps are as follows:

1. preparation of working fluid

1.1 intermediate solution: mu.L of the mother liquor from 10mM sample or control was removed and diluted with 495. mu.L of methanol (Conc.: 100. mu.M, 99% MeOH).

1.2 working solution: 50. mu.L of the intermediate solution was taken out and diluted with 450. mu.L of 100mM potassium phosphate buffer (Conc.: 10. mu.M, 9.9% MeOH).

Preparation of NADPH cofactor

Weighing appropriate amount of NADPH powder, and adding MgCl₂(10mM) solution.

3. Liver microsome

3.1 liver microsome information

3.2, preparation: the microsomal working solution was prepared with an appropriate concentration using 100mM potassium phosphate buffer.

4. Stopping liquid

Glacial acetonitrile containing 100ng/mL Tolbutamide (Tolbutamide) and 100ng/mL Labetalol (Labetalol) as internal standards.

5. Procedure of experiment

5.1 Add 10. mu.L of sample or control working solution/well to all plates (T0, T5, T10, T20, T30, T60, NCF60) except blank;

5.2 Add 80. mu.L microsome working solution/well to all plates, incubate 10min at 37 ℃;

5.3 Add 10. mu.L of 100mM potassium phosphate buffer/well to NCF60, incubate at 37 ℃ and start timer 1;

5.4 after preheating, 10. mu.L of NADPH/well was added to each plate to start the reaction;

final concentration of each component in the incubation medium:

incubation at 5.537 ℃ and activation of timer 1;

5.6 stop the reaction by adding 300. mu.L/well;

5.7 shaking the system for about 10 min;

5.8 centrifugation (4000rpm) of the sample at 4 ℃ for 20 min;

5.9 taking 8 new 96-well plates, adding 300. mu.L HPLC water/well, then adding 100. mu.L supernatant, mixing and making LC/MS/MS.

4. Hepatocyte metabolism assay

And (3) experimental operation: the sample to be tested (concentration 10. mu.M) or the positive control (concentration 30. mu.M) was mixed with hepatocytes of various genera (cell density 1.0X 10)⁶cells/mL) were incubated at 37 ℃/5% CO 2/saturated humidity for 120 min. The sample was centrifuged after precipitating the protein with 0.1% formic acid in acetonitrile at a ratio of 1:2, and the supernatant was dried with nitrogen. After blowing dry, redissolve with 200. mu.L of a 10% acetonitrile/water solution (containing 0.1% formic acid). 15 μ L of each sample was introduced into an LC-MS instrument system for analysis.

Possible metabolites detected by LXY-Cl in human hepatocytes

Potential metabolites of IB-33 detected in human hepatocytes

Based on the above results of hepatic microsome stability and hepatocyte metabolism, it was shown that the metabolic stability of the fluorine-containing substituted compound was significantly improved.

Claims

1. A compound of the general formula:

preferably, R₁Selected from: C14-C18 alkyl substituted by 1,3, 5, 7, 9, 11, 13 or 15 fluorine atoms;

more preferably, R₁Selected from: c₁₆FH₃₂-、C₁₄F₇H₂₂-、C₁₅F₉H₂₂-、C₁₆F₁₁H₂₂-、C₁₇F₁₃H₂₂-；

R₂Selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl;

preferably, R₂Selected from: hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl;

more preferably, R₂Selected from: hydrogen, methyl, isopropyl, cyclopropyl;

R₃is selected from

1)

Wherein Z₁，Z₂，Z₃，Z₄，Z₅Each independently selected from:

(1) hydrogen, fluorine, chlorine, bromine, iodine, nitro, cyano, amino, hydroxyl, hydroxycarbonyl, methoxyformyl, ethoxyformyl, N-propoxycarbonyl, isopropoxyformyl, carbamoyl, N-methylcarbonyl, N-ethylformyl, N-N-propylformyl, N-isopropylformyl, N-cyclopropylformyl, N-N-butylformyl, N-isobutylformyl, N-tert-butylformyl, N-cyclobutylformyl, N-N-pentylformyl, N-isopentylcarbonyl, N-cyclopentylcarbonyl, N-N-hexylformyl, N-isohexylcarbonyl, N-cyclohexylformyl, N, N-dimethylformyl, N, N-diethylformyl, N, N-di-N-propylformyl, N, N-diisopropylformyl, cyclopropylcarbamoyl, cyclobutylaminoformyl, cyclopentylaminoformyl, cyclohexylcarbamoyl, 4-hydroxypiperidinoylpiperazinyl, piperazinoylmethyl, 4-N-methylpiperazinoylmethyl, 4-N-ethylpiperazinoylmethyl, 4-N-N-propylpiperazinoylmethyl, 4-N-isopropylpiperazinoylmethyl, methylsulfonyl, ethylsulfonyl, N-propylsulfonyl, isopropylsulfonyl, N-butylsulfonyl, isobutylsulfonyl, hydroxysulfonyl, aminosulfonyl, N-methylsulfonyl, N-ethylsulfonyl, N-N-propylsulfonyl, N-isopropylsulfonyl, N-cyclopropylsulfonyl, N-N-butylsulfonyl, n-isobutylsulfonyl, N-tert-butylsulfonyl, N-cyclobutylsulfonyl, N-N-pentylsulfonyl, N-isopentylsulfonyl, N-cyclopentylsulfonyl, N-N-hexylsulfonyl, N-isohexylsulfonyl, N-cyclohexylsulfonyl, N, N-dimethylsulfonyl, N, N-diethylsulfonyl, N, N-di-N-propylsulfonyl, N, N-diisopropylsulfonyl, cyclopropylaminosulfonyl, cyclobutylamine sulfonyl, cyclopentylamine sulfonyl, cyclohexylaminosulfonyl, 4-hydroxypiperidinylsulfonyl, piperazinylsulfonyl, 4-N-methylpiperazinylsulfonyl, 4-N-ethylpiperazinylsulfonyl, 4-N-N-propylpiperazinylsulfonyl, 4-N-isopropylpiperazinesulfonyl, carboxamido, acetamido, propionamido, N-butylamido, isobutyramido, cyclopropylcarboxamido, cyclobutylcarboxamido, cyclopentylcarboxamido, cyclohexylcarboxamido, methanesulfonamido, ethanesulfonamide, N-propanesulfonamide, isopropylsulfonamido, N-butylsulfonamido, isobutylsulfonamido;

Z₆selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl;

2)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

3)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

4)

wherein Z₂，Z₃，Z₄，Z₅The same as defined in 1) above;

preferably, R₃Is composed of

Z₆selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl; preferably Z₆Is hydrogen or methyl;

more preferably, R₃Is composed of

even more preferably, R₃Is composed of

X^-anion of pharmaceutically acceptable inorganic acid salt or organic acid salt; preferably, X^-Is chloride ion, bromide ion, iodide ion, sulfate ion, phosphate ion, maleate ion, fumarate ion, tartrate ion, palmitate ion, oxalate ion, citrate ion, succinate ion, methanesulfonate ion, benzenesulfonate ion, p-toluenesulfonate ion.

2. A compound according to claim 1 which is the following: :

Most preferably, R₁Selected from: n-C₁₆FH₃₂-、n-C₁₄F₇H₂₂-、n-C₁₅F₉H₂₂-、n-C₁₆F₁₁H₂₂-、n-C₁₇F₁₃H₂₂-；

R₂Selected from: hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl;

preferably R₂Selected from: hydrogen, C1-C3 alkyl, C3-C4 cycloalkyl;

more preferably R₂Selected from: hydrogen, methyl, isopropyl, cyclopropyl;

S₁selected from: 1 ' -halo, 1 ' -C1-C6 alkoxy (preferably 1 ' -C1-C3 alkoxy);

S₂selected from: 4 '-halo, 5' -halo;

preferably, S₁,S₂Each is as follows: 1 ' -halogen, 5 ' -halogen, or 1 ' -C1-C6 alkoxy (preferably 1 ' -C1-C3 alkoxy), 4 ' -halogen;

X^-anions of pharmaceutically acceptable inorganic acids and organic acids; preferably, X^-Is chloride ion, bromide ion, iodide ion, sulfate ion, phosphate ion, maleate ion, fumarate ion, tartrate ion, palmitate ion, oxalate ion, citrate ion, succinate ion, methanesulfonate ion, benzenesulfonate ion, p-toluenesulfonate ion;

3. A compound according to claim 1 or 2, selected from the following:

4. A process for preparing a compound as claimed in claim 3, wherein,

the process for preparing compounds IA-1 to IA-3 is:

the process for preparing the compounds IB-1 to IB-58 is:

5. A pharmaceutical composition comprising a compound of any one of claims 1-3, or a stereoisomer thereof, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and optionally a pharmaceutically acceptable excipient.

6. Use of a compound of any one of claims 1 to 3 or a stereoisomer thereof, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, or a pharmaceutical composition of claim 5, in the manufacture of a medicament for activating 5' -adenosine monophosphate-activated protein kinase (AMPK) activity.

7. Use of a compound of any one of claims 1 to 3 or a stereoisomer thereof, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, or a pharmaceutical composition of claim 5, for the manufacture of a medicament for decreasing fatty acid synthesis, a medicament for inhibiting triglyceride and cholesterol synthesis, a medicament for preventing and/or treating obesity and type II diabetes, a medicament for preventing and/or treating tumors, a medicament for preventing and/or treating parkinson's disease, a medicament for preventing and/or treating alzheimer's disease, or a medicament for prolonging the lifespan of a mammal.